Method of treating tobacco with catalase and hydrogen peroxide

ABSTRACT

A method of treating tobacco is described which involves the use of physical force to cause liquid containing catalase and hydrogen peroxide to permeate the interstices of the tobacco where they are reacted with each other in situ. Such reaction enhances properties of the tobacco, and in certain cases the volume of the tobacco is expanded. The physical force necessary to cause the treating material penetration may take the form of rollers, flat pressure means or continous expression means.

United States Patent 1 Rosen METHOD OF TREATING TOBACCO WITH CATALASEAND HYDROGEN PEROXIDE [75] Inventor: William E. Rosen, 2055 WisteriaLn., Lafayette Hills, Pa. 19444 [63] Continuation-impart of Ser. No.198,662, Dec. 20,

1971, abandoned.

[52] US. Cl. 131/140 P; 131/143 [51] Int. Cl. A241) 03/18; A24b 15/06[58] Field of Search 131/143, 17, 140; 99/135 [56] References CitedUNITED STATES PATENTS 2,344,106 3/1944 Reed 131/140 P 2,432,074 12/1947Jennings 117/111 R 3,006,767 10/1961 Huckabee 131/140 P UX June 17, 19756/1966 (16 la Burde 131/141 3,256,888 3,612,065 10/1971 Rosen 131/1433,710,803 1/1973 Johnson 131/140 P FOREIGN PATENTS OR APPLICATIONS993,077 5/1965 United Kingdom 131/142 A Primary Examiner-Melvin D. ReinAttorney, Agent, or Firm-Stephen E. Feldman [5 7 ABSTRACT A method oftreating tobacco is described which involves the use of physical forceto cause liquid containing catalase and hydrogen peroxide to permeatethe interstices of the tobacco where they are reacted with each other in'situ. Such reaction enhances properties of the tobacco, and in certaincases the volume of the tobacco is expanded.

The physical force necessary to cause the treating material penetrationmay take the form of rollers, flat pressure means or continousexpression means.

13 Claims, No Drawings METHOD OF TREATING TOBACCO WITH CATALASE ANDHYDROGEN PEROXIDE CROSS REFERENCE TO RELATED APPLICATIONS Thisapplication is a continuation-in-part of my copending application Ser.No. 198,662, filed Dec. 20, 1971, now abandoned.

BACKGROUND OF THE INVENTION 1. Field of the Invention This inventionrelates to a method of treating tobacco, and more particularly relatesto a method for improving the quality of tobacco and increasing itsusefulness in the manufacture of smoking products.

2. Description of the Prior Art In my US. Pat. No. 3,612,065, thedisclosure of which is incorporated herein by reference and made a parthereof, I have described a method of treating tobacco wherein a givenamount of catalase provided on said tobacco is reacted with a givenamount of hydrogen peroxide in contact therewith, the mutualdecomposition of catalase and hydrogen peroxide being effective toincrease the volume of the tobacco up to percent. Other prior artmethods of treating tobacco to increase the volume thereof require theuse of expensive solvents and complex solvent recovery systems. Somechemical solvents may affect the tobacco taste or leave a toxic residue.9

Tobacco quality is determined by various objective and subjectivefactors, including the following:

1. Geographical source of the tobacco.

2. The portion of the tobacco plant used to form the tobacco product.

3. The moisture content of the tobacco.

4. The resistance to crushing of the tobacco.

5. The flavor of the tobacco.

6. The aroma of the tobacco before and during smok- It is known that asa result of importing certain foreign tobaccos importers andmanufacturers are supplied with amounts of low quality tobacco,including both stems and lamina. These products may have a musty orammoniacal odor and a sharp, peppery biting taste.

It is also known that conventional tobacco curing and ageing methodsrequire from several months to several years in time in order toobtain'desirable changes in the appearance, taste and aroma of thetobacco, as well as the smoke produced therefrom.

The less valuable forms of tobacco include the stems, ribs and stalks ofthe plant. However, these forms may be utilized in certain proportionsfor the production of acceptable smoking products. For example, it iswell known that cigars are often made with a wrapper of one qualitytobacco, a binder of another quality tobacco and a filler of stillanother quality tobacco. Obviously, the more less valuable tobaccoproduct which can be incorporated into a highly rated tobacco product,the greater are the economies which may be expected. This same reasoningpertains to cigarettes since such product is filled with shreddedtobacco, which is much more economically obtained from forms of tobaccosuch as stems, ribs, stalks and scraps than from the more expensiveforms of tobacco.

The anatomical and chemical structure of tobacco may be exemplified bythat of tobacco stems. Tobacco stem is herbaceous, consisting of pith,primary xylem, vascular rays and cortex. It is poor in collenchyma andrich in parenchyma, no cork being present. The cells are rectangular andtheir walls are thickened by thedeposition of cellulose and pectincompounds. In cured stem all cells shrink together because of the rapidmoisture loss, thereby reducing intracellular and intercellular space.However, the cells contain a great number of pores which permit thepassage of gases.

It is accordingly an object of this invention to provide an improvedmethod for treating tobacco with catalas and hydrogen peroxide.

It is another object of this invention to provide a method for treatingtobacco more thoroughly than heretofore with agents for enhancingqualities of the tobacco.

It is another object of this invention to provide an improved method ofexpanding the volume of tobacco.

SUMMARY OF THE INVENTION The objects of this invention have beenachieved by the process of treating tobacco which comprises the steps ofl evenly distributing on the surface of the tobacco a liquid solutioncontaining catalase and a liquid solution containing hydrogen peroxideand (2) applying physical force to said tobacco to cause a portion ofthe hydrogen peroxide to permeate the interstices of the tobacco,whereby the catalase and hydrogen peroxide react on and within saidtobacco. The selection of appropriate amounts of catalase and hydrogenperoxide will cause these reatants to mutually decompose. Where thephysical force is removed from the tobacco prior to completion of thereaction of catalase and hydrogen peroxide, the gases produced by saidreaction act to expand the volume of the tobacco.

DESCRIPTION OF THE PREFERRED EMBODIMENTS The essential feature of theprocess of this invention is the carrying out of a reaction betweencatalase and hydrogen peroxide from within the interstices of. tobacco.By so doing, the effects of said reaction are brought to bear morethoroughly on the tobacco constituents than by any other process knownto me. By

this means, I have been able to obtain increased expansion of tobaccoand excellent flavor and aroma enhancement.

It will readily be apparent that the order of applying the catalase andhydrogen peroxide to the tobacco is not critical. A solution containingcatalase may first be applied, then a solution containing hydrogenperoxide, or vice versa. Also, the catalase and hydrogen peroxide may beapplied in the same solution, in which case a delayed catalase ispreferred. Moreover, the catalase and hydrogen peroxide, if applied fromseparate solutions may be forced into the interstices of the tobacco atthe same time, or one at a time.

The physical force may be applied to the wetted tobacco in manyconventionalways. such as by weightedflat plates, rollers, presses, orcontinuous expression devices. In a preferred embodiment of thisinvention,

any excess liquid resulting from the application of physical force tothe tobacco is prevented from again contacting the tobacco when theforce is removed.

Since thecatalase and hydrogen peroxide will mutually decomposeproportionateamounts of themselves with the production of nascentoxygen, it is readily apparent that the physical force must be appliedin a timely manner in order that the optimum effect of such reactionwithin the interstices of the tobacco is obtained. It is believed thatthe flavor enhancement obtained by the process of this invention is dueto both sterilizationof bacteria within the tobacco and chemical changesin the tobacco brought about by oxidation.

EXAMPLE 1 A lb. sample of Pennsylvania cigar filler tobacco was treatedby spraying an aqueous catalase (4 ml. catalase/l I. water) solutiononto the tobacco in an amount equal to 10 percent of the tobacco weight,and I r EXAMPLE 2 Example 1 was repeated, except that immediately afterthe spraying of the-hydrogen peroxide solution the tobacco was rolledwith a heavy roller for a few seconds. When the rolling was stopped, thetobacco was seen to visibly expand to more than the volume of thetreated tobacco from Example 1 and upon smoking showed considerableimprovement in flavor over the control with a mildness that wasdescribed as being neutral.

EXAMPLE 3 Poor quality Phillipine flue cured cigarette strips weretreated by spraying an aqueous catalase (4 ml. catalase/l I. water)solution onto the tobacco in an amount equal to 10 percent of thetobacco weight, and then spraying onto the tobacco a 35 percent hydrogenperoxide solution in an amount equal to 5 percent by weight of thetobacco. This tobacco was dried and smoked with no noticeable change inflavor over the control which flavor was called undesirable to theindustry by virtue of being overly sweet.

EXAMPLE 4 Example 3 was repeated, except that after the spraying of thecatalase solution the tobacco was rolled with a heavy roller for a fewseconds, and immediately after the spraying of the hydrogen peroxidesolution the tobacco was again rolled with a heavy roller for a fewseconds. Upon drying and smoking the overly sweet flavor had become moreneutral and the tobacco had greater expansion.

EXAMPLE 5 EXAMPLE 6 Example 5 was repeated except that after thecatalase spraying the stems were pressed by a flat weighted plate for afew seconds. This tobacco was dried and smoked, the flavor being definedas neutral and cigarette like. The tobacco had expanded more than theresultant product of Example 5.

. EXAMPLE 7 Shredded cigarette lamina of low quality is placed in thewide end of a funnel having a central shaft with a helical screw surfaceformed thereon which closely conforms to the funnel. As the shaft isturned the to bacco is forced through the funnel being alternatelycompressed and released from the funnel. Before being forced through thefunnel this tobacco is sprayed first with an aqueous catalase solution(4 ml. catalase/l I. water) in an amount equal to 10 percent byweight ofthe tobacco and then sprayed with a 12 percent hydrogen peroxide in anamount equal to 5 percent by weight of the tobacco. After being forcedthrough the expression device the tobacco was dried and smoked, having amild smooth taste. The moisture content after expression was less than22 percent.

EXAMPLE 8 I Ten lbs. of a cigar tobacco was prepared by cutting 4-5 inchstrips from theleaf. This tobacco was first sprayed with a 5 percentportion of 907 ml. of an aqueous catalase solution (3 ml. catalase/l 1.water), then sprayed with 907 ml. of a 6 percent hdyrogen peroxidesolution to wet the tobacco 20 percent above its original moisturecontent, then sprayed with the remaining catalase solution. The thussprayed tobacco was rolled with a heavy roller for a few seconds. Thetreated strips were bulked and placed in a curing room for 20 days,where the temperature was controlled to not more than 140 F. The curedtobacco had a mild smooth sweet flavor and the leaves had obviouslyexpanded.

EXAMPLE 9 10 lbs. of a cut blend of domestically grown lamina I fillerhaving a strong odor of ammonia and a sharp bite was obtained. Thistobacco was first steamed. Then a portion of 680 ml. of an aqueouscatalase solution (4 ml. catalase/l 1. water) was sprayed on thetobacco. Then 680 ml. of a 25 percent hydrogen peroxide solution wassprayed on the tobacco, followed by the remaining portion of thecatalase solution. Then the sprayed tobacco was pressed by a weightedfiat plate for several seconds, after which the tobacco was againsteamed. The moisture level of the tobacco was never more than 29percent during this treatment and it was easily dried. The resultinglamina when smoked had a smooth mild flavor and based on fillingcapacity showed an increased volume of percent.

EXAMPLE 10 lO-lbs. of whole stems were obtained. This tobacco wastreated by (l) steaming well, (2) spraying with a portion of 907 ml. ofan aqueous catalase solution (4 ml. catalase/1 1. water), (3) sprayingwith 907 ml. of 12 percent hydrogen peroxide, (4) spraying with theremainder of the catalase solution, (5) rolling the tobacco with a heavyroller for several seconds, (6) steaming well and (7) drying in a drier.When the thus treated tobacco was cut, rolled and smoked, it had a,flavor which was sweeter and milder than a control,-

' and the volume thereof had expanded three times.

EXAMPLE 1 1 101 lbs. of cut rolled burley stems was obtained. Thistobacco was treated by l steaming well. (2) sprayingwith 680 ml. of anaqueous catalase solution (5 ml. catalase/l l. water), (3) spraying with680 ml. of percent hydrogen peroxide solution, (4) rolling the tobaccowith a heavy roller for several seconds, (5) steaming well, and (6)drying. Moistures did not exceed 30 percent during the process and uponsmoking the flavor of the stems was mild and sweet. Expansion was 100percent.

EXAMPLE 12 Cut stems were treated by wetting to a 40 percent moisturelevel with a combined spray of 1 percent hydrogen peroxide solution anda 1 percent aqueous delayed catalase solution. The sprays were combinedabove the tobacco and allowed to fall thereon. After spraying thistobacco was rolled for several seconds with a heavy roller, and thendried. The tobacco had a filling capacity of 150 per cent of controltobacco. Cigarettes were then made from this tobacco.

The cigarettes were conditioned for 25 hours at 75 F. and 60% relativehumidity. Smoking was done in a laboratory conditioned to 75 F. and 60R.H. The smoking system consisted of the cigarette, a tared Cambridgefilter assembly and a smoking machine that produces a 35 ml. puff of 2second duration at a rate of one puff per minute. All cigarettes weresmoked to a butt length of 3 mm longer than the tipping paper. Fivecigarettes were smoked through each Cambridge filter and the resultscalculated and reported in terms of one cigarette.

Moisture was determined by gas chromatography according to a proceduredescribed in a paper entitled Determination of Moisture in TotalParticulate Matter by Schultz and Spears in Tobacco Science, Vol. X, pp.75-76 (1966).

As soon as the filter assembly had been weighed to determine totalparticulate matter, it was opened and the filter holder was then wipedwith one-fourth of a Cambridge filter pad and this added to the bottle.Ten ml. of dry dioxane-isopropanol 100:1) was measured from an automaticburette into the serum bottle and the stopper inserted. The bottle wasshaken for minutes on a Wrist Action shaker.

A sample of 5 microliters was withdrawn with a Hamilton syringe throughthe rubber serum cap and subjected to gas chromatography in an Aerograph90-p3 gas chromatograph. The moisture content of the particulate matterwas read from a calibration curve made by adding known amounts of waterto the solvent mixture. Cigarettes conditioned at 75 F. and R.H. weretreated in the same manner and the average value of the moisture contentsubtracted from the amounts found to get the net moisture content.

The balance of the solution for GlC moisture, plus the filter pad, wastransferred to a Griffith still and the nicotine determined by the usualprocedure of double distillation and reading at three wave lengths in anultraviolet spectrophotometer.

The results of these tests were as follows:

Total Particulate Average number of puffs for all groups was [0 Buttlength 27mm Average pressure drop 6.1 inches EXAMPLE l3 Separate lots ofcut rolled tobacco stems were treated by first spraying the tobaccothoroughly with a 3% by weight acqueous hydrogen peroxide solution andthen with an acqueous solution of catalase, both solutions being appliedat a rate of 250 ml. of solution per pound of tobacco. The treatedtobacco was then rolled for several seconds with a heavy roller and thendried. Each lot was blended with tobacco lamina to give lamina/25% cutrolled stems blends. Comparable blends of untreated cut rolled stems andlamina were prepared as a control, and all of the blends made intocigarette samples and tested as follows:

Table No. 1A

UNTREATED TREATED REQUIRED C.R.S. c/i C.R.S. 227 ch C.R.S. 130 c/iC.R.S. 227 ch Moisture content before treatment 7:- 18 18 Used H Oconcentration 70 3 5 l0 3 5 10 Total Moisture Content after H 0application Z 32:2 32: 2 32" 32i2 32:2 32:2

Total Moisture content after Enzyma application (finished treatment) /1461-2 46+..2 46fl 46:2 46fl 4612 Used Drying Temp. F. 14015 140: 1401140:5 1405 Moist. content after 48 hrs. conditioning at 76F. 687: R.H.7! 16.4 16.2 17.6 17.5 17.4 17.5 17.5 17.5

Table No. 1A Cntinued UNTREATED TREATED REQUIRED C.R.S. 130 c/i C.R.S.227 c/i C.R.S. 130 e/i C.R.S. 227 c/i Compression m/m 6.60 8.98 4.372.32 I 59 4.84 3.58 1.88

Column Height m/m 23.40 22.02 25.63 27.68 28.41 25.16 26.42 28.12

Used Quantity: g. o1C.R.S. 20 20 20 20 20 20 20 2O Expansion 7r 9.5 18.321.4 14.2 20 27.7

Table 1B UNTREATED TREATED C.R.S. 130 c/i C.R.S. 227 c/i C.R.S. 130 c/iC.R.S. 227 6/1 Moisture content before treatment 7: 18 18 Used H O:concentration "A 3 1 10 3 5 10 Total Moisture Content aftcr H O:application 71 32 32:2 32:2 32*; 32-- 32:2 Total Moist. content afterEnzyma application (finished treatment) 7( 46:2 46:2 46:2 46:2 46:2 46flUsed Drying Temp. F. 1405 140:5 140:5 140:5 140:5 140:5 Moisture contentafter 72 hrs. conditioning at 73- 657: RH. 71 12.50 12.80 14.0 13.6 13.613.9 13.7 13.8 Compression Height MM 8.24 9.27 4.40 2.52 1.04 4.26 3.482.36 Column Height MM 21.76 20.23 25.60 27.48 28.96 25.74 26.52 27.64Used Quantity of C.R.S. 1

15 15 15 15 15 15 15 Expansion 7: 17.6 23.3 33.1 27.2 31.1 36.6

Table 1C Cigarettes Blends Untreated Cigarettes Blends Treated Cont. 7:Untr. C.R.S. 130 c/i in Lamina only Cont. 2571 Untr. C.R.S. 227 c/i in75% Lamina only Cont. 25'71 Treat. C.R.S. 227 c/i in 757: Lamina onlyCont. 257r Treat C.R.S. c/i in 7571 Lamina only Moisture Content in CRSLAMINA CRS LAMlNA CRS LAMlNA CRS LAMlNA C.R.S. And Lamina at 18 16 18 1618 16 18 16 Blending time '/1 Used H O Concentration 7: 3 3

Blends Moist. Contents After 72 hrs. conditioning at 73F. 657r R.H. 14.314.4 14.6 14.4

Compression MM 5.98 5.84 2.92 2.96

Column Height MM 24.02 24.16 27.08 27.04

Used Quantity of Blend:

Expansion 7: 13 12 Table No. 2

Samples Cgt. Wt. Moisture Diameter Compression Filling ExpansionDescription g. m/m m/m Index g. 7

Control 130 c/i 1.1570 14.00 7.93 5.54 1.0381

+9 Sample 130 c/i Treated with 371 1.1568 14.56 7.93 5.59 0.9404

Control 227 c/i 1.1582 14.20 7.98 5.52 1.0413

+8 Sample 227 c/i Treated with 3'7: 1.1540 14.37 7.90 5.58 0.9563

Table No. 3A Prior to cigarette manufacture, tobacco from various leafpositions within each group was composited and Sample PD TPM NICOTINEuniformly blended. Cigarettes 85mm 1n length were Description mg mgprepared usmg 32 cuts/inch, Cigarettes were selected 5 having the samedraw resistance. 225213 5? '87 I Smoking Procedure and ChemicalExamination of the Treated with 3% 13.2 20.03 0.89 Smoke H202 V Control227 C '3 2157 [07 The cigarettes were moisture equlllbrated at 65% relSample 227 c/i ative hum1d1ty and C. The cigarettes prepared from Trcmd21-09 lo the hydgrogen peroxide-catalase process were selected to weigh835 mg and the cigarettes made from the con- Table No. 3B

Sample Cgt. wt. Moisture Cold Warm Collapsing Description g. CompressionCompression lndex m/m m/m m/m Control 130 c/i 1.1570 14.00 4.47 4.471.07 Sample I c/i Treated with 37:

Control 227 ch l.l582 14.20 5.58 4.50 Sample 227 c/i Treated with 3%H2O? 1.1540 14.37 5.52 4.42 1.10 Repeat Control 227 c/i 1.2400 14.3 5.424.35 1.0? Sample 227 c/i Treated with 37! When cigarettes from thetreated samples are smoked, a concrete mild smooth sensation can bedetected. This is surprising since cut rolled stems as a cigarettefiller is a very rich-in-cellulose component which contributes directlyto the organoleptic qualities of smoke, and which when added to leaftobacco normally gave the smoke a harsh quality readily detected by thesmoker. Referring to Tables 1A, 1B and 1C, effective increases infilling values are shown for the treated lots of cut rolled stems andtheir blends with lamina. Results of blends made into machine madecigarettes are shown in Table 2. Referring to Table 3A, smoking testsshowed no significant differences on T.P.M. and nicotine between thetreated cigarettes and the controls.

Referring to Table 3B, no significant differences be- I tween thecollapsing indexes of the treated cigarettes and the controls werefound.

EXAMPLE 14 for several seconds with a heavy roller. When no fur therreaction was apparent, the tobacco of this group appeared bright incolor.

Following drying of the separate groups, samples were conditioned forhandling.

ventionally cured tobacco were selected to weigh 835 The two types ofcigarettes were smoked mechani cally on a Phipps and Bird machine usingCambridge Filters for trapping the particulate matter (puff volume 35ml, puff duration 2 sec., puff frequency 1 puff/min, butt length 23 mm).The determination of the amount of total particulate matter and itswater content were performed according to the Coresta standard methodNo. 10. The amount of nicotine was determined by the Coresta standardmethod No. 12. The determination of phenol in the particulate matterwhich accounts for 97 per cent of total smoke phenol value wasaccomplished using a method of Williamson modified by Carlson, which isbased on spectroscopic determination (480nm) after treatment of theseparated acidic material with diazotized p-nitroaniline. The results ofthese determinations are as follows:

Cut lamina was sprayed with a mixture of a 2% by weight acqueoushydrogen peroxide solution and a 1.5% by weight acqueous catalasesolution ((having a delay of 3 minutes) up to 20% by weight moisturelevel. This lamina was then rolled with a heavy roller for severalseconds and allowed to dry. The filling capacity of this treated laminawas 990 milliliters. Cigarettes were made from the untreated lamina and75% treated lamina 25% untreated lamina and 25% treated lamina 75%untreated lamina, and tested as follows: Smoking Conditions andCollection of Tar Twenty cigarettes were smoked for each set ofdeterminations. These were smoked to an approximate 23 35ml puff of 2seconds duration was taken once a minute on each cigarette. The smokewas collected at room temperature in 50ml. of vigorously stirredcyclohexane contained in a 250 ml. flask. Addition of the smoke beneaththe surface of the cyclohexane was avoided to eliminate a possiblepressure drop. Collection of smoke in this manner was very efficient.During the smoking of groups of cigarettes no significant amount ofparticulate matter deposited on a Cambridge filter which was placedbehind the collection flask in the smoking train.

After the smoke was collected, the cyclohexane solution was transferredto a separatory funnel. The collection flask, stirrer, and inlet tubewere rinsed with alternate portions of 2X5 ml. of 0.5 N sodium hydroxideand 3X5 m1 of cyclohexane. After removing the aqueous layer, thecyclohexane solution was extracted with 3X5 ml. of 0.5 N sodiumhydroxide followed by 1X5 ml. saturated NaCl sodium chloride. Theextracts were combined and backwashed with 2X5 ml. of cyclohexane. Thisbase extract was used to determine phenols. The combined cyclohexanesolution was extracted with 5X5 ml. of 0.5 N hydrochloric acid followedby 1X5 ml. of saturated sodium chloride. The extracts were combined andbackwashed with 2.5 ml of cyclohexane which was added to the maincyclohexane solution. This acid extract was used for the determinationof nicotine.

The cyclohexane solution was next extracted with 5 ml. of methanol water(4:1). After separation the methanol-water solution was backwashed with2.10 ml. and 4.5 ml. of cyclohexane which was added to the maincyclohexane solution. The cyclohexane solution was next extracted with2.10 ml. and 6.5 ml. of nitromethane. These nitromethane extracts werecombined and the solution concentrated to dryness using a rotaryevaporator (bath temp. 45C; press. mm.-). The residue was used for thedetermination of benzo-a-pyrene. Determination of Phenols The baseextract was steam distilled. Approximately 500 ml. of distillate wascollected and discarded. The

:ontents of the distillation flask were allowed to cool and thenacidified with ml. of 20% sulfuric acid and again steam distilled. Thedistillate (lml.) was acidi- Fied and extracted with 6.50 ml. ofpurifiedether. The ether extracts were combined and extracted with 2.5111. of saturated sodium bicarbonate solution. The ether solution wasdried over anhydrous sodium sulfate and concentrated to less than 1 ml.by distillation :hrough a 13 cm. Vigreux column at atmosphericpresmm.This concentrated ether solution was transferred :0 a 1 ml. volumetricflask and diluted to volume. This solution was used to determine phenolsby gas chroma- :ography, using a Barber-Coleman Selecta 5000 system vitha model 5043 electrometer and a flame ionization letector. A 2mXl/8 in.s.s. column packed with -120 mesh Parapak Q was used.

The operating parameters were: nitrogen carrier gas flow 25 ml./min; airand hydrogen gas flows, 300 ml. and 27 mL/rnin respectively; column,injector and detector temperatures, 230, 250, and 270C, respectively.The electrometer settings were: sensitivity, 10" amps and attenuation,2. An lnfotronics Digital Readout System, Model CRS-ll HSB, was used todetermine'peak areas. The settings on the integrator were: trackingrate,3 up-l0 down; threshold level, 0.05; trip, 4; slope sensitivity, 1; andfilter frequency, 1.

The concentration of Phenol, o-cresol, and mand/or p-cresol wasdetermined from the ratio of their individual peak areas to the area ofo-chlorophenol which served as an interanal standard. An aliquot of astandard solution containing 2 mg. of o-chlorophenol was added to thecyclohexane solution immediately after the smoke was collected. Therelative response and retention time of the phenols to o-chlorophenolwere determined from a standard solution containing phenol, o-, m-,p-cresol and o-chlorophenol.

Determination of Nicotine This procedure is a modification of thatdescribed by Ogg for the determination of nicotine in smoke particulatematter collected on a Cambridge Filter. The acid extract and 10 ml. of0.5 N hydrochloric acid were steam distilled until 300-400ml. ofdistillate was collected, which was discarded. After the acidic solutionin the distillation flask had been allowed to cool, 30 ml. of a 30%sodium hydroxide solution, saturated with sodium chloride, was added.The resulting solution was then steam distilled until approximately 400ml. of distillate was collected. In this second distillation thedistillate was collected underthe surface of 50 ml. of 0.5 Nhydrochloric acid contained in a 500 ml. volumetric flask. After thevolume of the distillate was adjusted to 500 ml. with distilled water, a25 ml. aliquot was diluted to 100 ml. with 0.05 N hydrochloric acid. Theabsorbance of this solution was read at 236 mu, 259 mu, and 282 muMilligrams of nicotine per cigarette, N, was calculated using theformula:

N A259 A A )/2/0.3213 x b,

where A A and A are the absorbances at the in- .dicated wavelengths andB is the pathlength'of the cell material. The eluant which contained thebenzo-apyrene and other aromatic hydrocarbons was concentrated todryness using a rotary evaporator (bath temp.

' 45C, press 30mm). The weight of this residue is reand concentrated toapproximately 0.1 ml. and chromatogrammed on a 20% acetylated cellulosethinv layer plate. After the plate was developed; inethanoltoluene-water (17:4:1), the benzo-apyrene band was transferred toa 15 ml. centrifuge tube and recovered by desorbing with 4X4 ml. ofmethanol. All methanol solutions were pooled. concentrated toapproximately 0.3 ml., and filtered through a small cotton plug into a'volumetric flask. After dilution to 2 or 5 ml. the fluorescencespectrum of the solution-was obtained, using an Aminco-BowmanSpectrofluorometer set at an excitation wavelength of 380 mu. Thefluore'scence'maximum at 405 mu. was read and the concentration ofbenzo-a-pyrene was determined with a standard curve. Determination ofTotal Particulate Matter Total particulate matter was determined fromfivev cigarette which was held in 8 Cambridge filter holder.

The burn temperature was recorded as the point of maximum recorderresponse as the combustion zone passed over the thermocouple junction.For each deterhydrogen peroxide solution and a 0.5% by weight aqueouscatalase (2 min. delay) solution was prepared. To

this solution was added an equal amount of ethanol.

Cut lamina was wetted thoroughly by a spray of this by drogenperoxide-catalase-alcohol solution. The wet lamina was then rolled forseveral seconds with a heavy Cigarettes were made from this treatedtobacco in varying proportions with respect to the control tobacco; Eachcigarette contained 1 gram of tobacco and, was'85 mm. in length. All ofthe cigarettes then were tested by smoking-in a constant volume smokingmachine using the following conditions: puff volume, 35 ml.; puffduration, 2 sec.; puff frequency, 1 per min.

Tests were then made of the tobacco smoke to determine the presence ofethane, ethylene, methanol, acetaldehyde, acetonitrile, acrolein andacetone in the smoke composition. Moisture determinations were performedby a modification of the Carl Fischer Method and with the Ohaus MoistureBalance, both of which gave comparable values. v

Temperature measurements were made using a Pt/Pt 13% R.l-l. thermocoupleof either 0.001 or 0.002 in. diameter. For combustion temperaturemeasurements, the thermocouple was inserted perpendicular to. the

mination 10 cigarettes were smoked, using a standard longitudinal axisof the cigarette at a point 15mm. from smoking cycle, and the valuesaveraged. the front end. The results of these tests were as follows:

' I Mol 7r Burn Temp. CO Methane Ethane Ethylene 100% P.'l.* 480C .870.02 .0008 .0001 75% P.T. 510 .98 0.05 .0012 .0005 50% P.T. 580 1.100.09 .0123 .0023. 0 25% P.T. 670 2.20 0.15 .0319 .014 puffed samplescomm control 849 3.00 0.43 .0805 .0389 75% 25% 40 Average 2/cig. (g)1.17 1.17 1.17 P mbacco Percent moisture 10.0 12.1 12.1 No. puff/20 cig.v 218 218 218 Average Burn temp (C) 497 668 878 Analysis of Cig. Smokeper cig. 153311525? I Z? i9? 533 Bum Awal- Acew- Nicotine (mg) Temp.Methanol dehyde nitrile lein Acetone $2321 2? (mg) 1 10092. P.T. 480C4.1 7.2 .09 .08 2.1 (mg) 7 8 15 75% P.T. 510 7.4 10.7 2.40 1.50 4.2cresol (Hg) 24 35 P.T. 580 10.2 14.6 3.82 2.81 8.0 Total phenols (Hg) 07 133 25% P.T. 670 18.3 24.2 6.91 per 00 Smoke control 849 36.1 74.014.32 Dry TPM (mg) .52 4.2 7.8 nicotine (mg) .53 .21 0.37 Benzo-a-pyrene(mg) .005 1.9. 3.0 Phenol (ug) 1; 2i @501 "8 Sampling Procedures 7. 9$5,E,,,;Z, f, 8% 2O 5 35 All analytlcal data were obtained on the fifthpuff of the cigarette. The puffing sequence was controlled by a presettimer (Dual-Trol), such that the interval between puffs was seconds andthe duration of the puff 2 seconds. Lightin the cigarette constitutedthe first 60 puff. A fresh Cambridge filter was used for each clgaretteto separate the particulate phase from the vapor phase. I Forcolorimetric analysis reagent solution was in- EXAMPLE 16 jected into adismantled collection flask with a syringe.

A mixture of equal parts of a 0.5% by weight aqueous The flask stopcockswere then closed, and the flask inserted into a wet ice-bath andconnected to the smoking machine manifold. At the end of the fourthpuff, the

sample flask was opened to the manifold and evacuated. After 45 seconds,the stopcock on the vacuum line was closed and after 60 seconds, a puffof smoke was automatically puffed into the flask during the secondinterval. The flask was then closed, removed, and its contentsthoroughly shaken prior to colorimetric analysis.

For gas chromatographic analysis aliquots of the smoke gas wereextracted from the manifold'by means of sampling values and admittedimmediately into the carrier gas stream of the chromatographic column.The volume of gas admitted to the column was regulated by the length ofloops on the sampling values.

Determinations were based on measurements of peak heights; peaks werecharacterized by retention time comparisons and chromatography withstandard mixtures of known compounds. Methane and CO 10 ft. and 0.25 in.o.d. column of 40-60 mesh molecular sieve 13x; flow rate 125 ml/min.ambient column temperature; thermal conductivity detection. Acrolein,acetonitrile, acetaldehyde, acetone and methanol-l ft. X 0.125 in. o.d.column of Polypak l 1; flow rate 45 ml./min., column temperatureprogrammed from 50 170 c. at 3.5/min. and from 170 225 C. at 2.8/min.,

hydrogen flame detection.

EXAMPLE 17 Cut lamina was wetted thoroughly by spraying with a mixtureof equal parts of 0.5% aqueous solution of hydrogen peroxide andisopropanol. This lamina was then rolled for several seconds with aheavy roller and then immersed in liquid nitrogen. Upon removal from theliquid nitrogen the lamina was wetted with a 0.5% aqueous solution ofcatalase and then rolled again for several seconds. This material wasthen vacuum dried for minutes. Measurement of the filling capacity ofthe thus dried tobacco showed 805 milliliters. Fifteen experimentalcigarettes were made using 35% of this treated tobacco and 65% cutlamina control. These experimental cigarettes were compared withcommercial Kool cigarettes by smoking each in lots of five cigaretteseach. The CO content of the mainstream smoke was determined Smokingconditions were as specified by the current Federal Trade Commissionregulation, 2-sec. 35-ml. puffs at 1 minute intervals in a laboratorycontrolled to 75%F. and 60% Rh. The following data were obtained:

Carbon Monoxide mg./liter Vol. 70

Kool commercial Lot 1 59 5.2 Lot 2 58 5.1 Lot 3 60 5 ,3 Average 59 5.2Experimental Lot 1 22 1 Lot 2 22 1 Lot 3 22 1 Average 22 l isms. Also inaccordance with the process of this invention, catalase and hydrogenperoxide are mutually decomposed, leaving no amount of either reactant.Peroxide-catalase treatment of milk for cheesemaking is known, as isperoxidase-catalase treatment of liquid egg products.

The fact that flavor of tobacco may be enhanced by the process of thisinvention is believed to be due to the fact that the extremely largeamount of nascent oxygen evolved upon decomposition of hydgrogenperoxide acts to oxidize substances within the tobacco as well as toexpand the tobacco. The resulting milder, smoother tastes are consideredto be the consequence of a breakdown of polysaccharide chains such asstarch and cellulose composed of glucose residues which are the mainconstituents of the cell walls of the tobacco leaf.

It will be apparent that there is no theoretical limit on the amounts ofhydrogen peroxide and catalase used, since the process of this inventionis shown to be effective at levels at least as low as 0.5% by weightconcentrations. Increased mutually decomposing amounts of catalase andhydrogen peroxide bring about greater recognized effects, such asexpansion, but the upper limits of catalase and hydrogen peroxide areonly economic.

It will moreover be appreciated that advantages accrue to the process ofthis invention when the hydrogen peroxide is made more concentrated froman aqueous solution while it is on and within the tobacco. Examples ofthe means for accomplishing this concentration are contacting theperoxide wetted tobacco with cold gaseous or liquid nitrogen, heatedair, ultraviolet radiation and/or vacuum drying.

Once the hydrogen peroxide has been concentrated it is then contacted bycatalase in its normal or delayed form inorder to puff the tobacco.Various inhibitors can be used to delay the catalase including citricacid and alcohol.

The advantages of concentrating the hydrogen peroxide are one of economyand efficiency. For instance, a lesser concentration of peroxide may beused to wet the tobacco, but when concentrated can double or triple instrength. The greater the strength of the peroxide, the greater theexpansion of the tobacco when contacted with catalase. Also concentratedperoxide will mix better with the catalase when contacted in that thereis less water present creating a maximum of the treating materials. Thishelps to eliminate excess moisture and insures quicker drying.Furthermore, the peroxide will penetrate down into the tobacco andconcentrate in the cells so that when catalase hits the tobacco surfacethe reaction will spread down into the tobacco expanding it to itsmaximum.

The method of applying the nitrogen and alcohol is by spraying orimmersion or other feasible methods or by passing an air stream over thetobacco as in the case of heated air.

It is also possible although not a preferred method to first treat thetobacco with nitrogen gas or liquid, alcohol or heated air. The hydrogenperoxide may then be applied. It will then concentrate although not asefficiently. The tobacco may also be treated by a series of treatmentsof either the nitrogen liquid or gas, alcohol, heated air, ultravioletlight, or vacuum drying. For instance, the tobacco may be sprayed withliquid nitrogen, then soaked in hydrogen peroxide, then contacted withcatalase and then heated air passed over it.

The tobacco may also be treated with hydrogen peroxide by spraying,immersed in nitrogen and contacted with catalase and then vacuum dried.

The tobacco may be sprayed with hydrogen peroxide, vacuum dried toconcentrate, and then contacted with catalase.

The tobacco may also be treated with a mixture of hydrogen peroxide anddelayed catalase placed on the tobacco with alcohol as a carrier. Thistobacco will then be contacted with heated air under 140 to concentratethe peroxide and volatilize the alcohol. The catalase already present inthe mixture will then react with the concentrated peroxide.

The tobacco may also be treated with hydrgen peroxide and alcohol andsprayed with liquid nitrogen to concentrate, contacted with catalase andthen vacuum dried.

Another method of treatment is to freeze the hydrogen peroxide on thetobacco and then contact it with liquid nitrogen to stabilize andconcentrate the hydrogen peroxide. The tobacco will then be contactedwith catalase to reactivate the hydrogen peroxide and then dried withheated air.

The alcohols used may be of the type that is nontoxic for use on tobaccosuch as ethanol, isopropanol and butanol.

While I have disclosed the use of the process of this invention withrespect to various naturally occurring forms of tobacco, such as leaf,stems and ribs, it will be readilly apparent that a finished productsuch as recon stituted tobacco sheet may likewise be treated inaccordance with the process of this invention to achieve the desirableadvantages thereof.

EXAMPLE 18 A finished homogenized tobacco sheet is sprayed with equalparts of a 27 per cent by weight aqueous hydrogen peroxide solution andan aqueous catalase (4 ml catalase/1 water) solution in order toincrease the moisture content of the sheet by 20 per cent of its weight.The wet sheet is immediately pressed with a heavy roller in order tocause the solutions to penetrate interstices of the sheet. The sheet wasthen dried. After drying the sheet was found to be more porous thanoriginally and a cigarette incorporating the shredded sheet was judgedto be milder than a cigarette incorporating untreated sheet.

One advantage gained by treating reconstituted tobacco sheet by theprocess of this invention is the sufficient modification of the tobaccoflavor to enable a smoking article to be produced utilizing solelyreconstituted tobacco sheet as filler. This is an important industrialfactor since the amounts of reconstituted tobacco sheet utilized incommercial American cigarettes rose to a level of per cent of the totalfiller in 1964.

Additionally, many forms of reconstituted tobacco sheet are formed withhigh densities. By use of the process of this invention, finishedreconstituted tobacco sheet may be provided at a lower density thanheretofore allowing more air penetration of the tobacco article andtherefore lower burn temperature. it has also been observed that lowerdensity sheet may become stronger with respect to the liklihood oftearing when used to wrap or bind cigars or cigarettes. Moreover, suchproducts have increased hygroscopicity, leading to products with longershelf lives and stability.

Having thus described my invention, I claim:

1. A method of treating tobacco to improve its flavor and expand thetobacco, comprising the steps of substantially evenly distributing onthe surface of the tobacco catalase and a liquid solution containinghydrogen peroxide, the amounts of catalase and hydrogen peroxide beingsufficient to mutually decompose each other, and before significantreaction between these two components obtaining, applying physical forcein the form of pressure means selected from the group comprising rollermeans, flat pressure means, and continuous expression means to saidtobacco to cause catalase ahd hydrogen peroxide to permeate theinterstices of the tobacco, whereby the catalase and hydrogen peroxidereact in situ on and within the tobacco.

2. The method in accordance with claim 1, wherein the physical force isremoved from the tobacco prior to the completion of the reaction ofcatalase and hydrogen peroxide, thereby allowing volumetric expansion ofthe tobacco.

3. The method in accordance with claim 1, wherein a catalase solution isfirst applied and then a hydrogen peroxide solution is applied.

4. The method in accordance with claim 1, wherein a hydrogen peroxidesolution is first applied and then a catalase solution is applied.

5. The method in accordance with claim 1, wherein step l comprisesapplying a liquid solution containing both catalase and hydrogenperoxide on the surface of the tobacco.

6. The method in accordance with claim 5, wherein the catalase isdelayed catalase.

7. The method in accordance with claim 1, wherein the tobacco is uncuredtobacco.

8. The method in accordance with claim 1, wherein the tobacco is curedtobacco.

9. The method in accordance with claim 1, wherein the tobacco is wholeleaf tobacco.

10. The method in accordance with claim 1, wherein the tobacco istobacco lamina.

11. The method in accordance with claim 1, wherein the tobacco istobacco cut ribs and stems.

12. The method in accordance with claim 1, wherein the tobacco isfinished reconstituted tobacco sheet.

13. A method according to claim I wherein during the treatment of thetobacco the moisture content of the tobacco does not exceed about 46weight percent of the tobacco.

1. A METHOD OF TREATING TOBACCO TO IMPROVE ITS FLAVOR AND EXPAND THE TOBACCO, COMPRISING THE STEPS OF SUBSTANTIALLY EVENLY DIDTRIBUTING ON THE SURFACE OF THE TOBACCO CATALASE AND A LIQUID SOLUTION CONTAINING HYDROGEN PEROXIDE, THE AMOUNTS OF CATALASE AND HYDROGEN PEROXIDE BEING SUFFICIENT TO MUTUALLY DECOMPOSE EACH OTHER, AND BEFORE SIGNIFICANT REACTION BETWEEN THESE TWO COMPONENTS OBTAINING, APPLYING PHYSICAL FORCE IN THE FORM OF PRESSURE MEANS SELECTED FROM THE GROUP COMPRISING ROLLER MEANS, FLAT PRESSURE MEANS, AND CONTINUOUS EXPRESSION MEANS TO SAID TOBACCO TO CAUSE CATALASE AND HYDROGEN PEROXIDE TO PERMEATE THE INTERSTICES OF THE TOBACCO, WHEREBY THE CATALASE AND HYDROGEN PEROXIDE REACT IN SITU ON AND WITHIN THE TOBACCO.
 2. The method in accordance with claim 1, wherein the physical force is removed from the tobacco prior to the completion of the reaction of catalase and hydrogen peroxide, thereby allowing volumetric expansion of the tobacco.
 3. The method in accordance with claim 1, wherein a catalase solution is first applied and then a hydrogen peroxide solution is applied.
 4. The method in accordance with claim 1, wherein a hydrogen peroxide solution is first applied and then a catalase solution is applied.
 5. The method in accordance with claim 1, wherein step (1) comprises applying a liquid solution containing both catalase and hydrogen peroxide on the surface of the tobacco.
 6. The method in accordance with claim 5, wherein the catalase is delayed catalase.
 7. The method in accordance with claim 1, wherein the tobacco is uncured tobacco.
 8. The method in accordance with claim 1, wherein the tobacco is cured tobacco.
 9. The method in accordance with claim 1, wherein the tobacco is whole leaf tobacco.
 10. The method in accordance with claim 1, wherein the tobacco is tobacco lamina.
 11. The method in accordance with claim 1, wherein the tobacco is tobacco cut ribs and stems.
 12. The method in accordance with claim 1, wherein the tobacco is finished reconstituted tobacco sheet.
 13. A method according to claim 1 wherein during the treatment of the tobacco the moisture content of the tobacco does not exceed about 46 weight percent of the tobacco. 